Page 35 - 《中国药房》2022年13期

·药学研究·

Legumain酶和线粒体双级靶向去氢骆驼蓬碱脂质体的制备及其

体外特性评价 Δ

伊帕尔古丽·阿皮孜 ，贺宏吉，王昭志，李喆喆，卡迪热娅·艾克拉木，白静雅，王 梅（新疆医科大学药学院，乌
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鲁木齐 830017）

中图分类号 R944 文献标志码 A 文章编号 1001-0408（2022）13-1565-08
DOI 10.6039/j.issn.1001-0408.2022.13.06

摘 要 目的 制备Legumain酶和线粒体双级靶向去氢骆驼蓬碱（HM）脂质体（KA@HM-LPS），并对其制剂学特性、体外抗肿瘤
作用及生物相容性进行初步评价。方法 首先，筛选KA@HM-LPS的制备方法和均质化方法，并对制备的脂质体进行表征。其次，
分别测定 KA@HM-LPS 的血清稳定性、体外释放率、溶血百分数，以及空白脂质体作用下的细胞存活率。最后，分别测定
KA@HM-LPS 作用下的细胞存活率、线粒体靶向性和对肝癌细胞迁移和侵袭的抑制作用。结果 选择薄膜分散法制备
KA@HM-LPS，其包封率为（90.50±0.62）％；选择挤出法作为KA@HM-LPS的均质化方法。所制备的KA@HM-LPS粒径、多分散
系数、Zeta电位分别为（211.40±11.67）nm、0.316±0.014和（－14.20±0.49）mV。在37 ℃、10％胎牛血清中，12 h后KA@HM-LPS
的粒径基本稳定；KA@HM-LPS在20％血浆中的体外释放曲线符合Weibull分布，具有缓释效果；当HM质量浓度为160 μg/mL时，
KA@HM-LPS的溶血百分数为（4.23±0.19）％，远小于游离HM，具有安全性。当空白脂质体的质量浓度达400 μg/mL时，LO2细胞
的存活率为（94.40±6.12）％，载体生物相容性较好。体外细胞实验结果显示，KA@HM-LPS 对 Legumain 酶过表达的肝癌细胞
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LGMN -SK-Hep-1的抑制作用显著高于对正常肝癌细胞SK-Hep-1的抑制作用；且与SK-Hep-1细胞相比，LGMN -SK-Hep-1细胞
对载体的摄取效率更高；KA@HM-LPS可以更明显地抑制LGMN -SK-Hep-1细胞的迁移和侵袭。结论 成功制备KA@HM-LPS；
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该脂质体可以有效抑制Legumain酶过表达的肝癌细胞的迁移和侵袭，并提高HM的血液相容性。
关键词 去氢骆驼蓬碱；线粒体；Legumain酶；脂质体
Preparation of Legumain enzyme and mitochondrial double-stage targeted harmine liposome and
evaluation of its in vitro characterization
Ipargul·Hafiz，HE Hongji，WANG Zhaozhi，LI Zhezhe，Kadirya·Akram，BAI Jingya，WANG Mei（College of

Pharmacy，Xinjiang Medical University，Urumqi 830017，China）
ABSTRACT OBJECTIVE To prepare Legumain enzyme and mitochondrial double-stage targeted harmine （HM） liposome
（KA@HM-LPS）and preliminary evaluate its pharmaceutical properties，in vitro antitumor effect and biocompatibility. METHODS
Firstly，the preparation and homogenization methods of KA@HM-LPS was screened，and prepared liposomes were characterized.
Secondly，the serum stability，in vitro release rate，hemolysis percentage of KA@HM-LPS and cell survival rate under KA@BLPS
were determines respectively. Finally，the cell surivival rate，mitochondrial targeting and inhibitory effects on cell migration and
invasion of KA@HM-LPS were determined. RESULTS KA@HM-LPS was prepared by the thin-film dispersion method，with
encapsulation efficiency of （90.50 ± 0.62）％ . The extrusion moulding method was selected as homogenization method of
KA@HM-LPS. The particle size，polydispersity index，and Zeta potential of KA@HM-LPS were（211.40±11.67）nm，0.316±
0.014 and（－14.20±0.49）mV，respectively. In 37 ℃，10％ FBS，the particle size of KA@HM-LPS kept stable after 12 h. In
vitro release curve of KA@HM-LPS in 20％ plasma conformed to Weibull distribution and had the property of sustained release.
When HM concentration was 160 μg/mL，the hemolysis percentage of KA@HM-LPS was（4.23±0.19）％，which was much
lower than that of free HM，with safety. When the mass concentration of KA@BLPS reaches 400 μg/mL，the survival rate of LO2
cells was （94.40 ± 6.12）％ ，and the biocompatibility was
Δ 基金项目 国家自然科学基金资助项目（No.81760637）；新疆维 good. Cell test results in vitro showed that，inhibitory effect of
吾尔自治区天山英才计划第三期资助项目；新疆天然活性组分和释药
KA@HM-LPS on liver cancer cells with overexpression of
技术重点实验室项目（No.XJDX1713）；新疆医科大学大学生创新训练
Legumain enzyme （LGMN -SK-Hep-1） was significantly
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计划项目（No.CX2021048）
higher than that of normal liver cancer cells SK-Hep-1；
＊第一作者 硕士研究生。研究方向：药物新剂型及药物传输系
compared with SK-Hep-1， LGMN -SK-Hep-1 cells had a
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统。E-mail：ipargull@qq.com
# 通信作者 教授，博士生导师，博士。研究方向：药物新剂型及药 higher uptake efficiency of the liposome；KA@HM-LPS could
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物传输系统。E-mail：wm630@163.com significantly inhibit the migration and invasion of LGMN -
中国药房 2022年第33卷第13期 China Pharmacy 2022 Vol. 33 No. 13 ·1565 ·